CML8 and GAD4 function in (Z)–3–hexenol–mediated defense by regulating γ–aminobutyric acid accumulation in Arabidopsis

(Z)–3–hexenol, a small gaseous molecule, is produced in plants under biotic stress and induces defense responses in neighboring plants. However, little is known about how (Z)–3–hexenol induces plant defense–related signaling. In this study, we uncovered how (Z)–3–hexenol treatment enhances plant resistance to insect attacks by increasing γ–aminobutyric acid (GABA) contents in Arabidopsis leaves. First, (Z)–3–hexenol increases the intracellular content of calcium as secondary messenger in Arabidopsis leaf mesophyll cells. Both intracellular and extracellular calcium stores regulate changes in calcium content. Then, CML8 and GAD4 transmit calcium signaling to affect (Z)–3–hexenol induced GABA content and plant resistance. Herein, CML8 interaction with GAD4 was examined via yeast two–hybrid assays, firefly luciferase complementation imaging, and GST pull–down assays. These results indicate that (Z)–3–hexenol treatment increased the GABA contents in Arabidopsis leaves based on CML8 and GAD4, thus increasing plant resistance to the insect Plutella xylostella. This study revealed the mechanism of activating plant insect defense induced by (Z)–3–hexenol, which guides the study of volatiles as biological pest control.